Dimeric collagen hybridizing peptides and methods of using

Abstract

Disclosed are peptide conjugates comprising an active agent, a spacer moiety, and a dimeric collagen hybridizing peptide comprising a first and second collagen hybridizing peptide, a linker; and a branch point, wherein the first and second collagen hybridizing peptides comprise the sequence of at least (GXY)n, wherein G is glycine, wherein X and Y are any amino acid, and wherein n is any number between 3 and 12. Also disclosed are methods of detecting denatured collagen in a sample comprising contacting a composition comprising any one of the disclosed peptide conjugates to a sample, wherein the active agent comprises a therapeutic agent, and detecting the presence or absence of binding of the peptide conjugate to denatured collagen, the presence of binding indicating the presence of denatured collagen in the sample. Also disclosed are methods of treating a disease or injury involving collagen damage comprising administering to a subject having a disease or injury involving collagen damage any one of the disclosed peptide conjugates.

We claim: 
     
       1. A solid support comprising at least one peptide conjugate, wherein the at least one peptide conjugate comprises
 a) an active agent; 
 b) a spacer moiety; and 
 c) a dimeric collagen hybridizing peptide comprising
 i) a first and second collagen hybridizing peptide; 
 ii) a linker; and 
 iii) a branch point, 
 
 
       wherein the first and second collagen hybridizing peptides comprise the sequence of at least (GXY) n , wherein X and Y are any amino acid, and wherein n is any number between 3 and 12, 
       wherein the first and second collagen hybridizing peptides are capable of forming a triple helix with collagen. 
     
     
       2. The solid support of  claim 1 , wherein the solid support is attached to the peptide conjugate via an attachment point. 
     
     
       3. The solid support of  claim 2 , wherein the attachment point is an amino acid residue. 
     
     
       4. The solid support of  claim 3 , wherein the attachment point is a glycine residue. 
     
     
       5. The solid support of  claim 1 , wherein the solid support is resin, polymeric beads, agarose beads, nanotubes, nanoparticles, magnetic beads, gold nanoparticles, surface coated with gold, acrylamide, cellulose, nitrocellulose, glass, gold, polystyrene, polyethylene vinyl acetate, polypropylene, polymethacrylate, polyethylene, polyethylene oxide, glass, polysilicates, polycarbonates, teflon, fluorocarbons, nylon, silicon rubber, polyanhydrides, polyglycolic acid, polylactic acid, polyorthoesters, functionalized silane, polypropylfumerate, collagen, glycosaminoglycans, and polyamino acids or any polymeric surface. 
     
     
       6. The solid support of  claim 5 , wherein the solid support is a nanoparticle. 
     
     
       7. The solid support of  claim 6 , wherein the nanoparticle is gold coated. 
     
     
       8. The solid support of  claim 5 , wherein the solid support is an agarose bead. 
     
     
       9. The solid support of  claim 1 , wherein the solid support is a thin film, membrane, bead, bottle, dish, fiber, optical fiber, woven fiber, chip, compact disk, shaped polymer, metal, particle or microparticle. 
     
     
       10. The solid support of  claim 1 , wherein the first and second collagen hybridizing peptides of the dimeric collagen hybridizing peptide are identical. 
     
     
       11. The solid support of  claim 1 , wherein the first and second collagen hybridizing peptides of the dimeric collagen hybridizing peptide are different. 
     
     
       12. The solid support of  claim 1 , wherein the first and second collagen hybridizing peptides comprise the sequence of at least (GXY) n , wherein X is proline, modified proline, glutamic acid, or aspartic acid. 
     
     
       13. The solid support of  claim 1 , wherein the first and second collagen hybridizing peptides comprise the sequence of at least (GXY) n , wherein Y is a modified proline, lysine, or arginine. 
     
     
       14. The solid support of  claim 1 , wherein the first and second collagen hybridizing peptides comprise the sequence of at least (GXY) n , wherein a glycine is modified as an Aza-glycine. 
     
     
       15. The solid support of  claim 1 , wherein the linker of the dimeric collagen hybridizing peptide is between the collagen hybridizing peptides and the branch point. 
     
     
       16. The solid support of  claim 1 , wherein the dimeric collagen hybridizing peptide comprises at least two linkers. 
     
     
       17. The solid support of  claim 1 , wherein the linker and branch point of the dimeric collagen hybridizing peptide are on the C-terminal end of the first and second collagen hybridizing peptides. 
     
     
       18. The solid support of  claim 1 , wherein the linker and branch point of the dimeric collagen hybridizing peptide are on the N-terminal end of the first and second collagen hybridizing peptides. 
     
     
       19. The solid support of  claim 1 , wherein the linker of the dimeric collagen hybridizing peptide is one or more glycine residues, aminohexanoic acid, or polyethylene glycol (PEG). 
     
     
       20. The solid support of  claim 1 , wherein the branch point of the dimeric collagen hybridizing peptide attaches to a linker which is attached to the first collagen hybridizing peptide and to a linker which is attached to second collagen hybridizing peptide. 
     
     
       21. The solid support of  claim 1 , wherein the branch point of the dimeric collagen hybridizing peptide is a lysine residue. 
     
     
       22. The solid support of  claim 1 , wherein the dimeric peptide comprises the formula 
       
         
           
                 
               
                     
                 
                   (SEQ ID NO: 11) 
                 
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
             
                
               
               
                
                
                
               
            
           
         
       
     
     
       23. The solid support of  claim 1 , wherein the dimeric peptide comprises the formula 
       
         
           
                 
               
                     
                 
                   (SEQ ID NO: 51) 
                 
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
             
                
               
               
                
                
                
               
            
           
         
       
     
     
       24. A method of detecting denatured collagen in a sample comprising:
 a) contacting a sample comprising denatured collagen to the solid support of  claim 1 , wherein the active agent of the peptide conjugate comprises a detectable moiety, 
 b) detecting the presence or absence of binding of the peptide conjugate to denatured collagen in the sample, wherein the presence of binding indicates the presence of denatured collagen in the sample. 
 
     
     
       25. The method of  claim 24 , wherein the solid support is a resin, polymeric beads, agarose beads, nanotubes, nanoparticles, magnetic beads, gold nanoparticles, surface coated with gold, acrylamide, cellulose, nitrocellulose, glass, gold, polystyrene, polyethylene vinyl acetate, polypropylene, polymethacrylate, polyethylene, polyethylene oxide, glass, polysilicates, polycarbonates, teflon, fluorocarbons, nylon, silicon rubber, polyanhydrides, polyglycolic acid, polylactic acid, polyorthoesters, functionalized silane, polypropylfumerate, collagen, glycosaminoglycans, and polyamino acids or any polymeric surface. 
     
     
       26. The solid support of  claim 1 , wherein the active agent of the peptide conjugate is a detectable moiety. 
     
     
       27. The solid support of  claim 1 , wherein the spacer moiety of the peptide conjugate is between the active agent and the first or second collagen hybridizing peptide.